Document 1QrM7zb7Y1ZkxrLj2Yqgad9Ed

FOR DU PONT USE ONLY AR226-2962 Du Pont HLO 432-91 Study Title Skin Sensitisation Test with A n p f l H H n i n Guinea Pigs Study Completed On July 31, 1991 Contracting Laboratory Pharmakon Research International, Inc. P.0. Box 313 Faverly, Pennsylvania 18471 Author Susan Armondi for E. I. du Pont de Nemours and Company Haskell Laboratory for Toxicology and Industrial Medicine Elkton Road, P. 0. Box 50 Newark, Delaware 19714 Study Monitor Scott E. Loveless Medical Research No. Laboratory Project ID Haskell Laboratory Report No. 432-91 eonwanv Sanitfced. Dees noi copain T S C A CB Page 1 of 60 Du Pont HLO 432-91 PHARMAKON R ESEA RC H INTERNATIO NAL, INC. WAVERLY, PENNSYLVANIA 18471 PHONE (717) 586-2411 COMPLIANCE STATEMENT FAX (717) 586-3450 This study was conducted in compliance with the Principles of Good Laboratory Practices (GLP) as promulgated by the following regulatory agencies: U.S. Food and Drug Administration, as stated in the Federal Register, 21 CFR Part 58, Friday, September 4, 1987. U.S. Environmental Protection Agency as stated in the Federal Register, 40 CFR, Part 792, Thursday August 17, 1989. U.S. Environmental Protection Agency as stated in the Federal Register, 40 CFR, Part 160, Thursday, August 17, 1989. Organization for Economic Co-operation and Development Guidelines for Testing Chemicals (OECD), ISBN 92-64 12221-4, adopted by the council at its 535th meeting on 12th May, 1981. Study No.: PH 424-DU-001-91 "To the best of my knowledge, this study was conducted in accordance with applicable Good Laboratory Practice regulations;, there were no deviations from these regulations that impacted on study conclusions." 2 Material Tested: Medical Research No.: Haskell No.: Haskell Test Code No.: Pharmakon Study No.: Physical Form: Purity; Composition; GENERAL INFORMATION Test Material 18,790 PH 424-DU-001-91 Du Pont HLO 432-91 Synonyms: Other Codes; - Contaminants; Stability; Material Tested: Purity: Synonyms : Other Codes: In the absence of visible evidence to the contrary, the test material was assumed to be stable under the conditions of administration. Positive Control Material N-Phenyl-1,4-Phenylenediamine 9BZ 1,4-Benzenediamine p-phenylenediamine PPD Aldrich Chemical Co., Lot #05012MK r Aldrich Chemical Co., Catalog #24139-3 YCf'"ft f'Ri 3 GENERAL INFORMATION (CONT.) Du Pont HLO 432-91 CAS Registry No.: 101-54-2 Stability: Sponsor: In the absence of visible evidence to the contrary, the test material was assumed to be stable under the conditions of administration. Du Pont Chemicals E. I. du Pont de Nemours and Company Wilmington, Delaware Materials Submitted By: Pharmakon Notebook No.: E. J. Greenwood Du Pont Chemicals E. I. du Pont de Nemours arid Company Jackson Laboratory Deepwater, New Jersey Study Initiated - Completed: In-Life Phase Initiated - Completed: 2/22/91 - 7/31/91 4/29/91 - 6/7/91 There are 60 pages in this report. Distribution: K. D. Dastur (1) W. J. Brock (1) P. G. Gilby (1) E. J. Greenwood (1) N. C. Chromey/S. E. Loveless (1) A -4 c/As'jiciiy DU PONT CENTRAL RESEARCH AND DEVELOPMENT HASKELL LABORATORY FOR TOXICOLOGY AND INDUSTRIAL MEDICINE Du Pont HLO 432-91 July 31, 1991 E. J. Greenwood Du Pont Chemicals Jackson Laboratory Deepwater, New Jersey Skin Sensitization with) n Guinea Pigs Attached is a report from Pharmakon Research International, Inc. (PH 424-DU-001-91). Included in the report are test methods, results and conclusions. .. an intradermal induction concentration of I/o and c h & T l e n g ^ o r S f r a t i o n s of 50% and 5% in distilled water, was tested on the clipped intact skin of 5 male and 5 female guinea pigs. Appropriate positive, negative, and vehicle control groups were used during this study. During the challenge p h a s a j i ^ ^ ^ h e m a was observed in any test animals treated with either 50% or was expected, a positive sensitization response (slight to severe erythema) was observed in 9 out of 10 positive control animals. In contrast, no dermal responses were observed in any of the 5 negative or 5 vehicle control animals when challenged with the test material, positive control material, and/or vehicle. Under the conditions of this study contact hypersensitivity in guinea pigs. iid not produce delayed SEL/alw V Scott E. Loveless, Ph.D. Senior Research Toxicologist - 5- f lo w ^ vn TSC A CBt DU Pont HLO 432-91 PHARM AKON R ESEARCH INTERNATIO NAL, INC. WAVERLY, PENNSYLVANIA 18471 PHONE (717) 586-2411 FAX (717) 586-3450 Skin Sensitization Test with H-18,790 in Guinea Pigs PH 424-DU-001-91 ) Submitted to E. I. Du Pont De Nemours Newark, Delaware . /I c m n r L ^ Susan E. Armondi, LAT Study Director u h \ 9 I Date \j (j / V?-w<'ipC 'ci i i,,wl Du Pont HLO 432-91 Skin Sensitization Test with H-18,790 In Guinea Pias SUMMARY Test Article H-18,790 at an induction concentration of 1.0% (w/v) and challenge concentrations of 50% and 5.0% (w/v) in distilled water was tested on the clipped, intact skin of male and female guinea pigs. p-Phenylenediamine (PPD) as a 35% (w/v) suspension in distilled water was used to demonstrate the ability of the test system to detect a skin sensitizer (positive control group). Distilled water served as a vehicle control. During the primary irritation phase, no dermal irritation was observed in the vehicle control animals. Slight erythema was observed at 24 hours in one animal receiving the test article at the 50% concentration. No erythema was observed at the 5.0% concentration sites. No erythema was observed in the positive control animals receiving PPD at a 35% concentration. During the challenge phase, no to severe erythema was observed in the positive control group treated with p-phenylenediamine at a 35% concentration. No signs of erythema were observed in the test article-treated animals at the 50 or 5.0% concentration sites at 24 or 48 hours. No signs of erythema were observed in the vehicle control animals. No signs of erythema were observed in the negative control animals receiving the test article, vehicle or the positive control articles. Under the conditions of this study, H-18,790 did not produce delayed contact hypersensitivity in guinea pigs at 50 or 5.0% concentrations. Work by: Study Director: Susan E. Armondi, LAT Reviewed and Approved for Issue: Susan E. Armondi, LAT Study Director 7 Company Saniifeed. Does no? ce rtain T SC A GBf Du Pont HLO 432-91 QUALITY ASSURANCE UNIT STATEMENT Study N o .: PH 424-DU-001-91 Study Director: Susan E. Armondi This study was conducted in compliance with the Good Laboratory Practice Regulations. The Quality Assurance Unit conducted the inspections listed below and reported the results to the study director and to management on the dates indicated. The following inspections were performed: Interval Date In Life Phase April 29, 1991 Reporting Phase July 18. 1991 Date OAU Report Issued: To Study Director July 18, 1991 To Management July 18, 1991 8 `.I Du Pont HLO 432-91 INTRODUCTION The purpose of this study was to evaluate the potential of H-18,790 to produce delayed hypersensitivity or allergic reactions when applied to the skin of guinea pigs. Sensitization was defined as a significant score increase at challenge over the response observed after the primary application of the test material to the test guinea pigs, or the response observed in the negative controls. A significant score increase defined as a 2-or-more step increase (e.g., from 0 to 2, from 1 to 3, etc.) has been used at Haskell Laboratory as the criterion in this study protocol to identify compounds that are sensitizers. The sensitivity of the test system to detect chemical sensitizers was evaluated with pphenylenediamine. This study was conducted according to EPA Good Laboratory Practice Regulations as listed on page 2 of this report. Areas of noncompliance are documented in the study records. No deviations existed that significantly affected the validity of the study. MATERIAL AND METHODS A. Animal Husbandry Young adult male and female Duncan Hartley albino guinea pigs were received from BuckberG Lab Animals, Tomkins Cove, New York. The guinea pigs were housed singly in suspended, stainless steel, h " wire-mesh cages. Each guinea pig was assigned a unique identification card on which the animal number, study number, dose level and sex were recorded and affixed to the cage. Guinea pigs were ear tagged. Purina Certified Guinea Pig Chow1* #5026 and water were available ad libitum. Guinea-pigs were acclimated and observed for general health during a period of approximately five days. Animal rooms were maintained on a timer-controlled, 12-hour light/12-hour dark cycle. Environmental conditions of the rooms were targeted for a temperature of 22" 3C (66- 77F) and . relative humidity of 30 - 70%. Any excursions outside these ranges were of small magnitude and/or brief duration and did not adversely affect the validity of the study. B. Protocol A copy of the signed original protocol is attached as Appendix A. A preliminary range-finding test was conducted to estimate the primary irritation potential of the test material. The results of the range-finding study were used to select the exposure concentrations for the main study. The main sensitization study consisted of 3 phases: a primary irritation phase, an induction phase and a challenge phase. During each phase, skin responses were scored according to the system presented in Table I. During the study, body weights were recorded weekly. 9 Comoany Sanitized. Coes on* contain TSCA CPI Du Pont HLO 432-91 The topical range-finding test was conducted on 2 males and 1 female guinea pigs ranging in weight from 300 to 358 grams. Aliquots (approximately 0.05 mL) of 1.0%, 10%, 50% and 100% (w/v) suspensions of the test material in distilled water were applied and lightly rubbed onto four separate test sites on the clipped, intact skin of the back of each animal. The primary irritation phase was conducted in 10 guinea pigs (5 males and 5 females), weighing from 403 to 442 grams, by applying and lightly rubbing in 1 drop (approximately 0.05 mL) of 50% and 5.0% (w/v) suspensions of the test material in distilled water onto two separate sites of the clipped, intact skin of each animal. In addition, 10 positive control guinea pigs (5 males and 5 females), weighing from 401 to 451 grams, were treated by applying and lightly rubbing in 1 drop of a 35% (w/v) suspension of p-phenylenediamine in distilled water onto the shaved, intact left shoulder skin of each animal. Five vehicle control guinea pigs (2 males and 3 females), weighing from 401 to 439 grams, were treated with distilled water in the same manner at one site only. Dermal responses were scored approximately 24 and 48 hours after application of the test material. Two days after the primary dermal irritation phase, the induction phase of the study was initiated using the same 10 test guinea pigs in which primary irritation had been evaluated. Induction consisted of a series of 4 intradermal applications (1 each week) of 0.1 mL of the test material at a 1.0% (w/v) concentration in 0.9% saline. A virgin site on the back of each animal was used for each induction. The same induction procedure was followed for the 10 positive control guinea pigs using 0.1 ml of a 1.0% (w/v) solution of p-phenylenediamine in saline. The vehicle control animals were similarly treated with 0.1 ml saline. Skin responses were evaluated approximately 24 hours after each induction. , Two weeks after the last induction treatment, the test guinea pigs were challenged for sensitization by applying and lightly rubbing in 1 drop of 50% and 5.0% (w/v) suspensions of the test material in distilled water onto two separate sites of clipped, intact shoulder skin. The sites used during challenge were the same used on each animal during the primary dermal irritation phase, respectively. The positive control animals were challenged for sensitization by applying and lightly rubbing in 1 drop of a 35% (w/v) suspension of p-phenylenediamine distilled water onto two separate sites of clipped, intact skin. The vehicle control animals were treated at two sites with distilled water, in the same manner. Also, 3 male and 2 female previously untreated guinea pigs, weighing 410 to 438 grams at study initiation, were treated by applying 1 drop each of 50% and 5.0% (w/v) suspensions of the test material in distilled water onto two separate sites of clipped, intact skin and served as negative control animals. In O'.-cijS&ny 'auiiiEKi-. D i.cH .iw. 10 vm Du Pont HLO 432-91 addition, these animals received 0.05 ml of distilled water and 0.05 ml of p-phenylenediamine at 35%. These animals also served as negative control animals for the vehicle and positive control materials. C. Record Retention All raw data and the final report will be stored in the archives of Pharmakon Research International, Inc., Waverly, PA. RESULTS AND CONCLUSIONS Weekly body weights are presented in Appendix B. A copy of raw data is presented in Appendix C. In the topical range-finding test, no to mild erythema was observed. Based upon the results of the dose-range-finding study, 50% proved to be mildly irritating and as per protocol, the dose concentrations used for the primaryirritation phase were 50% and 5.0%. The dose concentration used for induction was 1% and as per protocol, the dose concentrations chosen for challenge were 50% and 5.0%. During the primary irritation phase, no signs of erythema were observed in the test article-treated animals at the 5.0% concentration site. Slight erythema was observed in one animal at the 50% concentration site at 24 hours. No erythema was observed in the positive control animals. No erythema was observed in the vehicle control animals receiving distilled water. Individual animal data are presented in Table II. During the induction phase, no signs of erythema were observed in the test article-treated animals. No to mild erythema with and without edema was observed in the positive-control animals. No signs of erythema were observed in the vehicle control animals. Individual animal data from the induction phase are presented in Table III. . Dermal responses observed during the challenge phase are summarized in the following table (page 12). Individual animal data are presented in Table IV. ACT/RP5 V Company Sanitized. Does not contain TSCA OB! 11 Du Pont HLO 432-91 Summary of Skin Responses: Challenge Phase Positive Control PPD Test Animals H-18.790 Responses 35% 24 hr 48 hr 50% 5.0% 24 hr 48 hr 24 hr 48 hr No erythema 1/10 1/10 10/10 10/10 10/10 10/10 Slight erythema 2/10 4/10 0/10 0/10 0/10 0/10 Mild erythema 5/10 3/10 0/10 0/10 0/10 0/10 Moderate erythema 1/10 1/10 0/10 0/10 0/10 0/10 Severe erythema with necrosis 1/10 1/10 0/10 0/10 0/10 0/10 Responses Negative: Control H-18 .790 PPD 50% 5. 0% 35% 24 hr 48 hr 24 hr 48 hr 24 hr 48 hr Vehicle Distilled Water 24 hr 48 hr No erythema 5/5 5/5 5/5 5/5 5/5 5/5 5/5 5/5 No erythema Vehicle Control 24 hr 48 hr 5/5 5/5 At challenge, H-18,790 did not produce delayed contact hypersensitivity at 50 or 5.0% concentrations. Individual animal data are presented in Table V. dsr y- 12 csw ft Du Pont H LO 432-91 TABLE I SCORING SYSTEM USED TO EVALUATE SKIN RESPONSES Score Skin Reaction ' 0 No erythema or Edema 1 Slight erythema (barely perceptible, usually nonconfluent) 2 Mild erythema (well defined, usually confluent) 3 Moderate erythema 4 Severe erythema (beet redness, with or without edema, eschar formations or necrosis) If edema, blanching, necrosis or eschar formation occurred, they were indicated using the following code: ED = Edema N = Necrosis - E = Eschar . B = Blanching SN = Superficial Necrosis 13 GUINEA PIG NUMBER 1186 1102 1103 1104 1105 1106 1107 1189 1190 1110 Du Pont HLO 432-91 TABLE II PRIMARY IRRITATION PHASE SKIN RESPONSES OBSERVED IN TEST GUINEA PIGS FOLLOWING TOPICAL EXPOSURE TO H-18,790 LEFT FRONT 50% 24 hr 48 hr 00 00 10 00 00 00 00 00 00 0 0. RIGHT FRONT 5. 0% 24 hr 48 hr 00 00 00 00 00 00 00 00 00 00 14 Du Pont HLO 432-91 TABLE II (Contadi PRIMARY IRRITATION PHASE SKIN RESPONSES OBSERVED IN VEHICLE CONTROL GUINEA PIGS FOLLOWING TOPICAL EXPOSURE TO DISTILLED WATER GUINEA PIG NUMBER LEFT FRONT Distilled Water 24 hr 48 hr 1111 00 1112 00 1113 1114 00 00 1191 00 Company Sanitized. Dess not contain TSCA CBi 15 Du Pont HLO 432-91 TABLE II fCont'd PRIMARY IRRITATION PHASE SKIN RESPONSES OBSERVED IN POSITIVE CONTROL GUINEA PIGS FOLLOWING TOPICAL EXPOSURE TO P-PHENYLENEDIAMINE GUINEA PIG NUMBER 1121 1122 1123 1187 1125 1193 1194 1128 1129 1195 - LEFT FRONT 35% 24 hr 48 hr 00 00 00 00 00 00 00 00 00 00 16 Du Pont HLO 432-91 TABLE III INDUCTION PHASE SKIN RESPONSES OBSERVED IN TEST GUINEA PIGS 24 HOURS FOLLOWING INTRADERMAL ADMINISTRATION OF 1.0% H-18.790 GUINEA PIG NUMBER 1186 1102 1103 1104 1105 1106 1107 1189 1190 1110 . Week 1 Dose #1 (LEFT) 0 0 0 0 0 0 0 0 0 0 Week 2 Dose #2 (RIGHT) 0 0 0 0 0 0 0 0 0 0 Week 3 Dose #3 (LEFT) 0 0 0 0 0 0 0 0 0 0 Week 4 Dose # 4 (RIGHT) 0 0 0 0 0 0 0 0 0 0 CfTWipMny Sanitized. Dees not contain T SC CB! 17 Du Pont HLO 432-91 TABLE III (continued) INDUCTION PHASE SKIN RESPONSES OBSERVED IN TEST GUINEA PIGS 24 HOURS FOLLOWING INTRADERMAL ADMINISTRATION OF 0.9% SALINE - GUINEA PIG NUMBER Week 1 Dose #1 (LEFT) Week 2 Dose #2 (RIGHT) Week3 Dose #3 (LEFT) Week 4 Dose #4 (RIGHT) 1111 1112 1113 1114 1191 0 0 00 0 0 00 0 0 00 0 0 00 0 0 00 f?tssnvr 18 Du Pont HLO 432-91 TABLE III (continued) INDUCTION PHASE SKIN RESPONSES OBSERVED IN TEST GUINEA PIGS 24 HOURS FOLLOWING INTRADERMAL ADMINISTRATION OF P-PHENYLENEDIAMINE GUINEA PIG NUMBER 1121 1122 1123 1187 1125 1193 1194 1128 1129 1195 ED = edema Week 1 Dose #1 (LEFT) 0 0 0 1 1 0 0 0 1 0 Week 2 Dose #2 (RIGHT) 2ED 2ED 1 1 1 1 1ED 1 1 1 Week 3 Dose #3 (LEFT) 0 0 1 1 1ED 0 1ED 1 2ED 0 Week 4 Dose #4 (RIGHT) 1 1 1 0 1 0 1 0 1 1 Company Sanitized. Does not contain TSCA CBi 19 GUINEA PIG NUMBER 1186 1102 1103 1104 1105 1106 1107 1189 1190 1110 Du Pont HLO 432-91 TABLE IV CHALLENGE PHASE SKIN RESPONSES OBSERVED IN TEST GUINEA PIGS FOLLOWING TOPICAL APPLICATION OF H-18.790 LEFT FRONT 50% 24 hr 48 hr 00 00 00 00 00 00 00 0 0. 00 00 RIGHT FRONT 5.0% 24 hr 48 hr 00 00 00 00 00 00 00 00 00 00 \ . -Uii 20 Du Pont HLO 432-91 TABLE IV (cont'd) CHALLENGE PHASE SKIN RESPONSES OBSERVED IN VEHICLE CONTROL GUINEA PIGS FOLLOWING TOPICAL APPLICATION OF DISTILLED WATER GUINEA PIG NUMBER LEFT FRONT Distilled water 24 hr 48 hr 1111 1112 1113 1114 1191 00 00 00 00 00 Com pany Sanitized. Does not cor.tain TSC A OBJ 21 GUINEA PIG NUMBER 1116 1117 1118 1119 1192 Du Pont HLO 432-91 TABLE IV ( Cont/d) CHALLENGE PHASE SKIN RESPONSES OBSERVED IN NEGATIVE CONTROL GUINEA PIGS FOLLOWING TOPICAL APPLICATION OF H-18,790 DISTILLED WATER AND P-PHENYLENEDIAMINE LEFT FRONT H-18 .790 50% 24 hr 48 hr 00 00 00 00 00 RIGHT FRONT H-18 .790 5.0 % 24 hr 48 hr 00 00 00 00 00 LEFT REAR Distilled Water 100% 24 hr 48 hr 00 00 00 00 00 RIGHT REAR PPD 35% 24 hr 48 hr 00 00 00 00 00 22 Du Pont HLO 432-91 TABLE IV (Cont'd CHALLENGE PHASE SKIN RESPONSES OBSERVED IN POSITIVE CONTROL GUINEA PIGS FOLLOWING TOPICAL APPLICATION OF P-PHENYLENEDIAMINE GUINEA PIG NUMBER 1121 1122 1123 1187 1125 1193 1194 1128 1129 1195 LEFT FRONT 35% 24 hr 48 hr 22 22 43 21 21 34 11 22 11 *0 0 Company Sanitized. Does nof contain TSCA C Si 23 Du Pont HLO 432-91 APPENDIX A PROTOCOL Du Pont HLO 432-91 1 P H A R M A K O N RESEARCH IN T E R N A T IO N A L . INC. W A V E R L Y , P E N N S Y L V A N IA 18471 Protocol - 424D . Skin Sensitization Test in Guinea Pigs PHONE (717) 586-2411 FAX (7171 586-3450 Sponsor; Testing Facility; Test Facility S.O.P. No.; Study No.; Purpose of the Study; Ownership of the Study; Study Monitor: Study Director: Q.A.U. Responsible Personnel: Dates of Performance: Good Laboratory Practices Statement; IACUC Statement; Tentative Date of Submission of Draft Report: E.I. duPont de Nemours Haskell Laboratory for Toxicology and Industrial Medicine Newark, DE 19714 Pharmakon Research International, Inc Waverly, Pennsylvania 18471 .' PH-424 To be assigned upon study initiation. D U "O P V This study is designed to evaluate the dermal sensitization potential of the test material xn guinea pigs. The sponsor owns the study. All raw data, analysis, and reports are the property of the sponsor. SUtf . PlvD- nolores-E-- Malek, Ph.D., E.I. duPont de Nemours Susan Armondi, LAT, Pharmakon Research International, Inc. . | Leslie J. Pinnell, M.S. Study will begin within one month of the receipt of the test chemical and authorized protocol. This study will be conducted following the U.S. Environmental Protection Agency Toxic Substances Control Act Good Laboratory Practice Standards (40 CFR Part 160) or Good Laboratory Practice Standards /an rpv Part 792) This protocol has been reviewed by the Institutional Animal Care and Use Committee (IACUC) and complies with acceptable standard animal welfare and humane care. Within three weeks following the completion of the in-life phase of the study. 080489 Company Sanitized. Does noi conan TSCA CB 25 Protocol - 424D^ Skin Sensitization Test in Guinea Pigs Du Pont HLO 432-91 Records Maintained: Raw Data: Record Retention: analytical Chemistry: All raw data, final reports, documentation and protocol will be maintained in the Pharmakon Central Files. Amendments to protocol Feed Lot Number Body weights, initial, weekly and final Compound preparation Volume administration Observed signs Observed skin reactivity Maintained in a Standard Pharmakon' Notebook All raw data and completed notebooks. Analysis and stability of the test article and test article/carrier mixture is the responsibility of the sponsor. If requested by the sponsor, Pharmakon Research International, Inc. will, through its subcontractor, conduct appropriate analytical analysis and will indicate the additional cost involved following receipt and evaluation of the appropriate analytical method. In the case where a satisfactory method is not provided, Pharmakon Research International, Inc. or its subcontractor, at additional cost to the sponsor, will develop appropriate methods. If the sponsor elects to analyze the test article/carrier mixtures, Pharmakon Research International, Inc. will collect the sample at appropriate designated intervals during the study and transmit the samples to the sponsor for analysis. TEST SYSTEM Species: Strain: Supplier: . (Source) : Sex: Guinea pig Hartley BuckberG Lab Animals, Tomkins Cove, New York or from any other O.S.D.A acceptable source. Male and Female (the sex used will be documented in the study records and final report) . Age: Weight at Initiation: Young Adult 400 - 650 grams 080489 3t ri:T 26 Cd: Du Pont HLO 432-91 Protocol - 424D 3 Skin Sensitization Test in Guinea Pigs No. on Study: Dose-Range-finaing: 3 Vehicle control: 5 (at the discretion of the Study Monitor, this group may be excluded from the study.) Negative control: 5 (if re-challenge is necessary, a second set of 5 naive, age-matched negative control animals will be used.) Test article: 10 Positive control: 10 (at the discretion of the Study Director after consultation with the Study Monitor, this group may be excluded from the study.) Method and Justification for Randomization: Treatment groups will be housed by vertical cage positioning, Any animals showing evidence of clinical signs during acclamation will be eliminated from randomization. Randomization is carried out by use of a computer generated random number table. Before treatment commences, all guinea pigs will be weighed, ranked according to body weight and assigned to treatment groups using a table of random numbers so that each treatment group will have a similar distribution according to body weight. Guinea pigs beyond the extremes of the body weight range will not be assigned to treatment groups. If there is a statistically significant difference in mean body weight between any two groups of the same sex after allocation to the treatment groups, substitution from spare animals will be used to correct the situation. Acclimation Period: Minimum: five (5) days WJ System of Cages marked with an animal group number, sex and Identification: treatment group. Guinea pigs are ear tagged. HUSBANDRY Research Facility Registration: Animal Rooms: U.S.D.A. Registration No. 23-107 under the Animal Welfare Act 74: SC 2131 et seq. Separate isolation by test system. Light cycle - 12 hours light, 12 hours dark. Temperature/Humidity - Maintained at a temperature of 22#C 3C and a humidity of 30 to 70%. Housing: Sanitization: Food: Guinea pigs are housed individually in stainless steel *j" wire mesh cages. Size in accordance with the "Guide for the Care and Use of Laboratory Animals" of the Institute of Laboratory Animal Resources, National Research Council. Waste material will be removed daily. Cages and feeders will be sanitized every two weeks. Purina Certified Guinea Pig Chow #5026 or acceptable substitute will be provided ad libitum. Feeders are 080489 Company Sanitized. Does not contain TSCA CB\ 27 Protocol - 424D^ Skin Sensitization Test in Guinea Pigs Food Analysis: designed to reduce soiling, bridging, and scattering. The potential effects of dietary contaminants have been considered, and based on the manufacturer's data, contaminant levels are believed to be within acceptable limits. There are no contaminants expected to be present in the dietary material known to be capable of interfering with the purpose or conduct of the study. Water: Water Analysis: Availability - fresh tap water, ad libitum. Water is monitored for contaminants at periodic intervals according to Standard Operating Procedure ) methods Rationale for Test System: This study is intended to provide information on the health hazards likely to arise from exposure to the test material by the dermal route; skin contact is a possible worker and consumer exposure route. Rationale for Dose Selection: Based upon the.results of the dose-range finding study and/or at the sponsor's request. Justification for Test System: According to EPA Health Effect Test Guidelines EPA 560/6-82-001. The guinea pig has historically been used in the evaluation of chemicals for dermal KN sensitization. Compound Preparation: Liquids Liquids will be applied as received except when specific instructions request dilution or mixing. The use of a vehicle for-dilution will be described in the study records and final report. Solids . The test material will be pulverized if appropriate. The test material may be moistened with distilled water or a suitable vehicle to ensure good contact with the skin. When a vehicle is used, the influence of the vehicle on penetration of the skin by the test substance will be considered. Absorption of the Test Material: Semi-Solids Extremely viscous liquids may be heated to approximately 50C to dosing, with the approval of the Study Monitor. The procedure used will be documented in the study records and included in the final report. For the purpose of this study, all of the test . material applied to the test sites is assumed to be absorbed by the test system. All calculations and the evaluation of effects will be based on the applied 28 DU pont HLO 432-91 Protocol - 424D 5 Skin Sensitization Test in Guinea Pigs Vehicle: Volume Administration: Route of Administration: Rationale for Route of Adminis tration: No. and Description of Animals per Dose Group: Method of Study Performance: The choice of vehicle varies, depending on the nature of the test substance. Commonly used vehicles are distilled water, saline, ethanol:distilled water (80:20, v/v), dimethyl phthalate, acetone:dimethyl phthalate (1:9, v/v) or propylene glycol. The vehicle used will be documented in the study records and the final report. Approximately 0.05 ml/dose . _ The test and control materials will be applied topically and/or injected intradermally, to the clipped, intact shoulder skin. Skin contact is a possible worker and consumer exposure route. Dose-range-finding: 3 Vehicle control: 5 (at the discretion of the Study Director, this group may be excluded from the study). Negative control: 5 (if re-challenge is necessary, a second set of 5 naive, age-matched negative control animals will be used). Test Article: 10 . Positive control: 10 (at the discretion of the Study Director, after consultation with the Study Monitor, this group may be excluded from the study). Dose-range-finding Study Prior to initiation of the main study, a dose-range-finding study will be performed. In order to evaluate the irritation potential of the test material, 3 animals will be subjected to preliminary studies as follows: ' Animals will be closely clipped over the back and sides. The test material concentrations will be based on the solubility of the test compound (solids will be tested at the maximum soluble concentration and at 2 or 3 lower concentrations). Liquids will be tested at 100% (as received or as prepared per the Sponsor's instructions) and at 2 or 3 lower concentrations. Each guinea pig may be dosed with up to 4 different concentrations at different sites (1 concentration/ site). In some cases where the solid is not soluble in a common solvent it will be tested as received by topical application. One drop (approximately 0.05 ml) of each concentration will be applied directly on the appropriate test site and rubbed gently with the end of the dosing syringe to cover an area of approximately 25 mm in diameter. 080489 Company Sanitised. Does not contain S C a C`b,: 29 Du Pont HLO 432-91 Protocol - 424D'1' Skin Sensitization Test in Guinea Pigs Observations for signs of dermal irritation will be recorded at 24 and 48 hours after application. At each observation, all treated sites will be scored according to Table I. 4 U Voltar sl'flU Primary Irritation Phase For the test guinea pigs there will be 2 sites PeJf animal in the area between the shoulder and the middle, of the back. One site coitcentration-whir-h --is~j o nihYfa>-Han -mTldl-y ^iitoting luwi-brih+fK, -lb. as determined from the dose-range-finding study. The second site will receive one-tenth of that concentration. Each site will receive i drop (approximately 0.05 ml) of the appropriate concentration, rubbed gently into the clipped s k m with the end of the dosing syringe to cover an area approximately 25 mm in diameter. The vehicle contro guinea pigs, if used, will be treated at x site as above using the vehicle only. The positive control guinea pigs will be treated at 1 site with the positive control. The negative control guinea pig will remain untreated until the challenge phase. Observations for signs of dermal irritation will be recorded at 24. and 48 hours after application. Sensitization Phase (Induction) She first induction application will be made after th 18-hour dermal observation is made for the primary .rritation phase. If the test material is soluble in mpropriate vehicle, e.g., 0.9% saline or dimethy jhthlate, induction will be performed by intradermal Injection. If the test material is insoluble in the :ommonly used vehicles a topical induction procedure The hair at the appropriate application site will e clipped on the day prior to each application or, if a topical induction procedure is employed, at least weekly during induction. After clipping, the area to be treated may be delineated for the convenience of and at the discretion of the study director or the experimenter. The hair will be clipped as needed during the study for evaluation of the dermal irritation. A total of 4 intradermal injections will be made, 1 per week at 1 week intervals. A virgin site will be used for each injection. A 0.1 ml aliquot of a 1% solution of the test material in an acceptable solvent (e.g., 0.9% saline, dimethyl phthalate, etc.) will e administered by intradermal injection to the clipped sacral area of each test guinea pig. If distilled water is used as a vehicle in the primary irritation phase, 0.9% saline will be used for the intradermal injections. The vehicle control guinea pigs will be 080489 30 Du Pont HLO 432-91 Protocol - 424D1 Skin Sensitization Test in Guinea Pigs Method Validation/ Positive Control; Evaluation of Dermal Response: treated as above using the vehicle only. The positive control guinea pigs will be treated as above using the positive control material. Topical Administration A total of 9 applications will be made to the sacral area, 3 applications per week for 3 weeks. One drop (approximately 0.05 ml) of a minimally irritating concentration of the test material in the acceptable solvent (e.g., dimethyl phthalate, 0.9% saline, etc. will be applied directly to the skin and then gently rubbed in with the end of the syringe, to cover an area approximately 25 mm in diameter on each test guinea pig. The vehicle control guinea pigs will receive similar treatments using the vehicle only. The positive control quinea pigs will receive similar treatments using the positive control article. Challenge Phase _. Approximately 14 days after the last induction treatment, the challenge treatment will be administered. The test material will be administered in the same manner as in the primary irritation phase, treating with the same concentrations at the same sites. Following the same procedure the negative control animals will be treated with the same concentrations of the test material, and may be treated with the vehicle and positive control material (up to 4 separate test sites). The vehicle control animals will be treated with the vehicle only. The positive control animals will receive the positive control article. If the test results are conclusive and the test material has either clearly caused allergic sensitization or is not a sensitizer, the test is terminated and the guinea pigs are euthanized. . If the reactions are less than conclusive, a second challenge (rechallenge) may be performed approximately 1 week after the initial challenge using the.same test guinea pigs and additional naive control guinea pigs. The vehicle and positive control animals may not be rechallenged. . The procedures and test system will be evaluated with a known dermal sensitizer to confirm the validity of the test. Evaluation of the test system with a known dermal sensitizer may not be conducted concurrently with every study, but testing with a known dermal sensitizer will be performed periodically. Dermal evaluations will be made between 23-25 hours and/or 47-49 hours following treatment, but the data will be reported as the 24- or 48-hour evaluations. 080489 Company Sanitized. Does not contain TSCA CBi 31 Du Pont HLO 432-91 Protocol - 424D1 8 Skin Sensitization Test in Guinea Pigs Interpretation of Results: . Sensitization Rating: Intervals a. Primary Irritation Test - Dermal evaluations will be made approximately 24 and 48 hours after the test material application. ' b. Induction - Dermal evaluations will be made approximately 24 hours after each induction, treatment. c. Challenge and Rechallenge (if required) - Dermal evaluations will be made approximately 24 and 48 hours after the test material applications. All test sites will be graded on a. scale of 0 to 4 as follows: TABLE I Score Skin.Reaction 0 Ho erythema or edema 1 Slight erythema (barely perceptible, usually nonconfluent) Mild erythema (well defined, usually confluent) Moderate erythema Severe erythema (beet redness, with or without edema, eschar formation or necrosis) If edema, blanching, necrosis, superficial necrosis or eschar formation occur, they will also be indicated using the following code: ED = Edema N ** Necrosis E *= Eschar B *= Blanching SN * Superficial Necrosis Sensitization is defined as a 2-or-more step increase (e.g., from 0 to 2, frcm 1 to 3, etc.) in dermal irritation scores in the test animals over those observed in the negative controls at challenge. When the test and negative control animals have similar dermal irritation scores, the test material will not be considered a sensitizer. The rating of a sensitizer is as follows: Ratio of Responsers Degree of Sensitization 1 to 2/10 Weak Sensitizer 3 to 4/10 Mild Sensitizer 080489 32 DU Pont HLO 432-91 Protocol - 424D1 9 Skin Sensitization Test in Guinea Pigs Ratio of Responsers 5 to 6/10 Degree of Sensitization Moderate Senitizer 7 and above Strong Senitizer This rating scale is not fixed rigidly. Scientific evaluation of the data and judgement will be used to decide whether a material will be classified as a weak, mild, etc. sensitizer. Necropsy: Pathological evaluations of test animals will not ordinarily be performed. Exceptions will be noted in the study records and final report. Euthanasia: Animals that surive the test period will be euthanized according to the Testing Facility's Euthanasia SOP and incinerated. Statistical Analysis: Statistical analyses of data will not be performed, unless requested by the sponsor. Rechallenge: ' The sponsor will be notified of animals that are considered sensitized. Verbal instructions for rechallenge will be given by the Sponsor, followed by written confirmation. All test article animals, along with an additional group of four naive guinea pigs, can be rechallenged six (6) days after primary challenge, but not before. The negative and positive ' control groups will not be rechallenged. POSITIVE CONTROL ARTICLE . Positive Control Article: N-Phenyl-1,4 phenylenediamine (PPD) Supplier (Source): . Aldrich Chemical Company, Milwaukee, Wisconsin Special Handling Instructions: Standard precautions Analysis of Purity: How Supplied: Rationale for Positive Control Article: The purity is the responsibility of the manufacturer. 100 gm bottle As per sponsor's request. ATXPL4 080489 Company Sanitized. D^es not c o r Fain TSCA CBF 33 Protocol - 424D^ 10 Skin Sensitization Test in Guinea Pigs Du Pont HLO 432-91 Test Article (Name or Code); Chemical Abstract No. or Code No. : Analysis of Purity/Stability; Carrier Mixtures: Analysis of the purity and stability of the test article is the responsibility of the sponsor. Analysis for stability, uniformity.and correctness of concentration of the test article in the carrier is the responsibility of the sponsor. ___ Return Test Article Carrier Mixtures to the Sponsor y* Dispose of Test Article Carrier Mixtures Person to whom carrier mixtures should be sent: V l f i - ___________ '------------ Shipping Instructions; AMENDMENTS ;^ c c J | L w J 2 X fAKf r ^ . ,'V-eA Date Date ATXPL4 a ? APPROVAL OF PROTOCOlLi T i Study Monitor Alh'Xl,Y&___________ Study Director Q j \ r Y Y ) < J ) n c l ^ not civsusi 34 Du Pont HLO 432-91 xi APPENDIX A Test Article Information Identification: Test Article (Name or Code) : 13 1 q O Lot or Sample No: Physical Description Purity: Expiration Dati Density/Specific Gravity Solubility (check one) : Water, lAcetone Ethanol 1 J o TM Uil DMSO Other (please specify)_ Chemical Classification: Flammable_ Corrosive Other II Storage Information: Material Storage (check one): Room Temperature X ; Refrigerator Freezer_______ ; Other (specify) ____ III Handling Information: Known Hazards: Precautions: Routine use of protective clothing includes laboratory coats, latex gloves, dust masks, and safety glasses. Other recommended precautions A v * tPf.u. Al/oll Q n - W w . K In Case of Emergency Related to ffcii * ZtfiJ this substance, contact: Sc*-*- e . Love-Uy (person) of (company/division) at 3 * 7 - ~ 4 ri~ 4 ^ ^ (phone number) IV Disposition: All materials will be returned to the sponsor three months following submission of the final report to the sponsor. Person and address to whom test articles are to be returned. fj{(\- Name: Address : VI Signature : ATXPL4 Date :____ /9/ Company Sanitized. Does not contain TSC CB Du Pont HLO 432-91 APPENDIX B WEEKLY BODY WEIGHTS (a ) 36 :.(.onpr.r,v 3l Du Pont HLO 432-91 WEEKLY BODY WEIGHTS (q) Animal Number Sex Initial Week II Week III Week IV Week V Week VI Test Group 1186 M 428 488 574 630 692 746 1102 M 441 515 592 561 721 777 1103 M 428 486 545 611 685 717 1104 M 442 ' 505 575 631 685 727 1105 M 412 463 503 544 593 638 1106 F 403 460 511 558 599 628 1107 F 409 435 503 531 573 619 1189 F 414 441 481 523 547 590 1190 F 406 446 495 530 530 607 1110 F 432 476 518 578 607 654 Final 765 795 750 755 661 638 599 . 606 628 672 Company Sanitized. Does n o t contain TSCA CBi 37 Du Pont H LO 432-91 Animal Number Sex Initial Positive Control Group 1121 1122 1123 1187 1125 1193 1194 1128 1129 1195 M 451 M 411 M 431 M 434 M 443 F 402 F 407 F 420 F 444 F 401 WEEKLY BODY WEIGHTS (cM Week II Week III Week IV 544 611 668 465 511 564 496 545 616 462 521 543 502 574 635 458 540 573 448 485 534 467 538 584 479 529 526 435 467 517 Week V 740 623 673 593 705 649 585 648 611 563 Week VI Final 804 830 669 680 737 736 612 604 741 780 676 722 615 638 678 651 629 626 593 613 >j 38 Du Pont HLO 432-91 Animal Number____Sex Initial Vehicle Control Group 1111 1112 1113 1114 1191 M 418 M 439 F 423 F 436 F 401 WEEKLY BODY WEIGHTS fer Week II Week III Week IV 488 548 596 503 580 570 436 486 497 453 496 529 454 480 516 Week V 647 638 497 558 540 Week VI Final 680 695 637 634 528 528 600 584 556 576 Company Sanitized. Does not contain T S C A C B 39 Du Pont HLO 432-91 Animal Number Sex Initial Negative Control 1116 1117 1118 1119 1192 M 438 M 414 M 410 F 411 F 427 WEEKLY BODY WEIGHTS (a Week II Week III Week IV 532 626 696 481 599 608 478 546 614 477 521 578 481 523 569 Week V 770 672 680 618 611 Week VI Final 834 840 726 722 706 731 658 676 650 659 40 DU Pont H LO 432-91 APPENDIX C COPY OF RAW DATA 41 Sempany SanFfized. Does m i confain TSCA CBt UU F o n x ilLiU 432-91 PHARMAKON RESEARCH INTERNATIONAL, INC. /S'*Sponsor :-- Skin Sensitization Test in Guinea Pias Dose-Range-Finding Study rf.. fari J, N.B.# I W O Page ,? X I Purpose:^ ^ tfl ftfi/K "' e, oS Stud^ No---_ W STM S , ^^ ^ Method : rcfi f h t f / Date of Initiation: 7/ Orf/ _ Date of Termination: Test Article: // ~ / ^ f ----- -'t < 1.____ _ uDeescsrcipntpiounoJ n i ^ ^ ^ ^ ^ H Amount Received Date Submitted: Vehicle: -- I-I<rmnr\nwrl-- L'(,rI_-_-_-_-_-_-_- DDoossee LLeevveellss::! Q ;10 , 5 5 fini! ( 1 0 % Route of Administration: Animal P.0. 8: ^ O f ! r k l ^ _______________ / sPecies = flinec*- P, Food Lot t : O Q - ^ A f ^ No. of Animals- o-.n. Study ---- - Strain: Identification: sSex: Male ^)-- if/y. FFeemmaallee Scale8 fatgdZic j 2 { h ) \ [ f t \ d /) J Light Cycle Checked: Bh n tn--1 _ _ y/ Animals~ciipped: A* / _________ _ f Compound Preparation:-------/Qff>/e ^ O - O ^ ^ u / ^ U \ I ATABL3(1) instigator jM A fk k O u lA A n m c rrila ' Study Director Dat Date - bXv'j -J MCi O a fi: ^ Ci;H Du Pont HLO 432-91 PHARMAKON RESEARCH INTERNATIONAL, INC. N .B .f Skin Sensitization Test in Guinea Pigs Page Dose-Range-Finding Study In this test, both sides of-the. animal are shaved and exposed to four concentrations of the test material. The different concentrations are applied concurrently to four different test sites. A given concentration is applied to a different test site on each guinea pig so.as to minimize variation in . response due to afferent skin locations. The format for application of the test article is-described below:____ ________ - Figure A --- Figure B Site 1 =. f - 0 / . Site 2 = 0 % Site. 3 = $Cf/c Site 4 lCO/0 1-4: Test Sites Animal No. / Sex (T Test article Initial Tifflfe of Site and dose Wt. (gms] Dose 1 &o l-o. 10. S D G r r \ Ct), 13 S 300 PMuJ p p Site Site 23 O/ 0 o y io Site Final 4 Wt. (qms) 3 Jr/,, ~S75 / P/3 V/vh'l_ vl/thiiA ' 'Z / . 48 Hour Scores ^ ......... - ...... -........ O/ o \0 / n o fO o 1 V / tJ-hj ] U -- Tr^/estigator y?/ Date ATAI3L3 (2) Study D ire c to r Date Company Sanitized. Does not contain TSCA CBi 43 D U F o n t fi.LU 4 3 2 - 9 1 PHARMAKON RESEARCH INTERNATIONAL, INC. N.B.# /770 Page ft C Skin Sensitization Test in Guinea Pigs Titie : Stem's ;T/-s*Zs*`v / e s r 6 - v ^ e * '/eC Sponsor: -ZT. duP*jr<J<? ^Je^o^S '*r*'*} it cteS'f+'&f **< Purpose: -c<7-/-TVtf.-Vc p/gj ' 3c*ss *^ Study No.: ?//Ki-/-!) - crp/-?/ -v '^tC 7X Method : /fe/gyg. ?4> P/toTocoC Technical Initiation: V a ^ / d O' oh IVTechnical Termination: Test Article: H - / 1 7 f O ______________ Positive Control :/ty Lot Number: Description nM Lot Number :_ d r / r f r 1' Description: ifos'AfiM'- Vehicle : a /P^t <a A---m--o--u--n--t-----S--u--b--m--i--t- t--e--d---:------------------------------------------------- Receipt Date: 0 . - ' 7 t _________ Animal p.o. ft-.6()ii-e>4i'Tliii-Sou cyln4/6 Vehicle: J L Route of Administration : Toxicology Lab. No .: )/f / 0 Species: Weight Range: "P,c_________________ Strain: /?,<;7 7 e y ______ Food Lot ft: S a tflh A Z>l9//A DWlA/ C No. of Animals on Study: '//)Iptjjj/3 ^ y Scale ft: Sex: Male / S ' __ Female/^Light Cycle Checked :y/df/4//? / ? V f /fr/ ft Rnimals Clipped: H h r k , C & ' d v k t * f h k a s//v/?/& -'&feer " ' /' . 7 y'" 77 Format for Primary Irritation Induction and Challenge Phases 1-2 Primary Irritation and Challenge 3-6 Sensitization Phase (Induction) C ATABLE3 Dtudy D ire c to r 7fi Du Pont HLO 432-91 CJ Notebook # / Pa^e . S_____ ..PHARMAKON RESEARCH INTERNATIONAL, INC. Dermal Sensitization Study: Guinea Pig Maximization Test : ' .............. - ....... .GENERAL OBSERVATIONS . Study No.: M W * t r t ) f / - C D F y i Compound: /4 .Sponsor Date ~ " 1 -* ~ "AN = (appears normal) /fa/aU .............Initials JJ. f/d 4 'ri/K s /ff'fxC //2/~ Z/2<-//#T//9Y, f' ' ' y Z ZZ _ Z Z Z ZZ Z Z Z Z Company Sanifized. Does noi contain TSCA CB 45 / Du Pont HLO 432-91 m N.B. g PageS , 3 % < PHARMAKON RESEARCH INTERNATIONAL, INC. c Skin Sensitization Test in Guines Pigs Test Article Preparation Study N o .: J J ' i / P V ~ b < J - c r o / " ' ........ ........ Test Article: / '3 ' 7 ' f C ) .. .... Sponsor:'gr.Z~. J u r Z ^ T Je. /l/c./*nt>a<2> Primary Irritation Phase (Week I) ;: . - `/tV,%-__>__ - - - V, ?y / r^-<X(yr.or r-(~Ssif f t 2SP/H&1 c S j Z i f J J r c Induction (Week III):' & / , 0 <f o - 7 / W A n M < f / J a i f A & L m J 2 ^ ? .f % A a b fi ' f/ * n / jf* fr-^7 / & * * & d ffPfil S /PA tl)' T c & J ciM u d dfoAj'CLl(lfl.//jP/P /n A K i ? V I) ' u -JJ L -------- ----------------------------- --------------------------- 1 7 / 5 / 9 / 4 6 A T A B L 3 (3) `.v.if&ny '''er-&C,I..K/ 46 Du Pont HLO 432-91 N.B. ft t~]rj O c PHARMAKON RESEARCH INTERNATIONAL, INC. Skin Sensitization Test in Guinea Pigs Page Z Z Test Article Preparation (Continued) Study No.: f y - Y g . < / - > & - rp/- f / c C A T A B L 3 (4) Company Sanitized. Does not contain TSCA GSi 47 Du Pont HLO 432-91 Notebook 8 Page 8 PHARMAKON RESEARCH INTERNAIONAL, INC. : Skin Sensitization Test in Guinea Pia; .... -- Body Weights (grams)' -^Sponsor : T . h ,, Test Article: -n- -- - --~ Study Number: ,h_c-CO/ '/. Lot Number: ITBFTTTTnTJTT, Animal ' a' dumber Sex " .*<{o /,-=*7 - has - v f7 In itia l Week I Week I-I 441 - t- Week I I I Week IV :- S 'j - S?4 L 5 l Week v 0 > ^ ...... 'IV G ?<A 1 hn 7< 597 / ? 7/ ^ v r< > . - uh - 7 / 7 750 ho'l - 7 ym - >>5 - t t - '3 '/ - 6 e? 6 - - r p * 1 5 5 t Hi ? . . . 4 / . HlZ ......PN* - _ % s ... Hu> - - //l .... 42.... i? n7/ yQ 4 /4 -id- 460* .... 4 3 6 HU H iU >3 . tH 5 7 3 7 / .. --- 1 ...... - t s .... 7 3 H it 5P3 6 Hn w 630 6 5 / W /i --- ^6 /*9f ^6.J9f9! t?O ftL C o l <bS .!///.. 4 - 43'/ tf// 41U- . . \Tt 4S:*-Jbq-W ,, "5-/3V / /K MM to ']... tt Si-7/9/ e r? _ 0-3-7' _f&AtJ 6s7A -t--- -- - - -- L '\ ATABL2(5) ' -gn/g 6AAt 7 55te^ M !0 m c L -- 1^ l[ L 48 Du Pont HLO 432-91 Notebook Page ft PHARMAKON RESEARCH INTERNAIONAL, INC. Skin Sensitization Test in Guinea Pia?; Body Weights (grams) Sponsor : K.t-h>7T Test Article: jA - VfcfnmPS ________ Study Number : -Ol.v-nT:l - Ql Lot Number: f^pj- f) n / r f i J l L U knimal Initial umber 5ex Week I Week II Week III ^[M .lI .tW.L g Hit lli.Hi F 1n3 -Gt. .im! G tt Sa 42b 46 453 4% h91 'to/ 464 WO ^374, %:40%* Week IV Week V b Ll l & (*3f 414 ?7 06$ $61 6W ( yo </K&k>S*' 37w tfeek VI Final (rfO m t& t S s ff Lp 6-&/3%-b466/ S7<> w bFynAi\ur r (. - t? ^/38 633 if30> Lr?6 n io ?64 HP u n _ Mrl un -JiiS... fi0~? Hio 4u 4V 4lY 64> il 7 &! S` m U/4 (pib a rt ,n 16> 7M n.'o, 73U U67 61(e iiQa $ Ji l '`f'W77A(3u', 4YI 5-0:1! 633 'SZ'r^ 6L$ bti 0$^/ bnh! (Z> 6 6M-9/ w Ceffleany Sanitised. Does not contain TSC CBt 49 Du Pont HLO 432-91 Notebook # H10 Pcige # PHARMAKON RESEARCH INTERNAIONAL, INC. Skin Sensitization Test in Guinea Pias Body Weights (grams) Test Article: 4 _ftpCjr, Study Number:4^ ^ g 4-hu .rp|-Cj( Lot Number: rini-drtnhrshU. Animal Initial Bumber Sex Week I Week II Week III Week IV Week V Week VI Final ^VhoA-AV -IIal j _ 46i m jS L 4il -1193 j S L 4SI , utn 434 4% <tua -1194 S - 4 4 3 $ u9S $ 4o9 4 5 ? $ 467 44? ha $ Hb 4ir7 -h e fi J L 4 4 4 4 7 ? - life 4 _ 461 434 ~!n%3r~ 64/ Lshf n 4o fo S 730 s i St4 U 3 U S ? c,9n S/6 SqI (*1Z 493 4X1 I S Lp4) GcS 14 U 3S lo 6 14/ iw S/6. 373 /S/b> 4*6 634 4 7 f Lp/S yCGM^So I .3 f 6 i{ ( S f u>n CYJS'-'r/ I Sfr* fall L & ? W6;e7rI6QL 407 is t * <S S S 6V> ?%-(<$ sonh/ ----6=^-- f f lf i Cp)'3. S nzSji&, 4 # -- ATABL2(5) . <-*t i- -tw-ry.! i r u-t.,-t5 `..v;. 50 r Sponsor; C .T ,/Yi4hwT ]V. Test Article: fl <?;fo rv PHARMAKON RESEARCH INTERNATIONAL, INC. Individual Animal Scores During Primary Irritation Phase I Study;Number; -9 / . _________ Notebook # 1 ^ 7 ^ Lot Numbel: kU 1 'age ' I M Animal Dose Cone. Dose Cone. Time o f ' ;24 ; Time o f . i i '.Time of No . / su 1L 0 3 //>3 H(.4 l/X lux,, Ubi ntf it% H . 1 to 4Sex Left Shoulder Right Shoulder Dose Hours ?Reading ! T i r.dOPfT' / ,7 5? S' 'V iT : ^ `>l^ ! \` \ S' V , . cs. \ 0 ; j ; i 1/ a : ii i ;0 o <r 0 V e ... \ !O 1 f -- % ~T~ 4 4 1 ^ ;-------------- --.: &Ov.-- ;-- i i oo . A0 i . i ; t 1 1 V . oo 0 0 : : i/ / 06 CQMY, M^lf--6*3oV:t 1Hours !'Reading !* ;0 Q_ L i M4 , !'1 ! \ i- 0 o oD. a Q_ A 0 0 0 ! 1' 1i : -1 /* :;_: !ii ! j ii i i! , /A i\ 1; 1 ? : ... oa\ 00: 1k s/ R.5a->0iam"is, i 1 _- Initials 1: ' i 1 i 1 ! s I ! 1 : i 1 1 ji ; - i -1 -4 --1 1 _. i}i_;___;:_ . _ i _ii_:-------- ; 1_Lti U: !_____ j1i i i _J.__i'_____ : . i1 \! M o u l d y b'Axg/if'^KotUcM. ATABL3 (9) Du Pont HLO 432-91 Company Sanitized. oea not contain TSCA o 03 Ln NJ 6 rm ^ PHARMAKON RESEARCH INTERNATIONAL, INC. Individual Animal Scores During Primary Irritation Phase Sponsor: p T hb WfoOueS T e s t A r t i c l e : _____ jj - __________ Study N u m b e r -JMi CC\I - 9 / ________ Lot Number:__ t r l A ______________ __ ____ Notebook # H 7 Page # 091 Anim al No. INI /11 o ! Iii^ ukJ 1i9 i Dose Cone. Dose Cone. / Tim e o f 24 Sex L e f t S h o u ld er R ig h t Shouldex' Dose Hours tff 6 `7 9R. O-+ -- i& r.v u m O O V/ / o / ./ o / r.z z P m o -- s / ------- - H--XH? -iItti.A, Tim e o f R eading' /< cX> /-- \ / ' I* *-*""l*A 48 Hour's Tim e o f R e a d in q 5.- __ / o\ ____ ; JL___ 1_ o ; \/ oS T `7ZZX7/ / , __ _ VPfhlU-W , Cohti L Ulta f f inn f f 111$ f f ufi 4 11$s 4- / / / // / -------- f f ------- " \ i i 1 * : 1 Du Pont HLO 432-91 in itia ls A T A B L 3 (9) PHARMAKON RESEARCH l'ERNATIONAL, INC. Individual Animal Scores During Primary Irritation Phase Sponsor: r r r W t h a t j v Test A rtic le :_ - - V ^ ____________ Study Lot Number: M _________ :-- Notebook . IV7 Page #__ , J ) - in UJ DU Pont HLO 432-91 Company Sanitized. Dees not conta..' TSCA G8i ATABL3 (9) * lr n ri in ( M O S o ;r-: <.; Spon sor: P .r. Je AJe T o s t A r t i c l e : / / --/ 2 ? 9 __________ PHARMAKON RESEARCH INTERN ATIO N AL, I N C .. I n d iv id u a l Anim al S core s D ring In d u c tio n S tu dy Number: 0-0/ - 9 /________ L o t Num ber:____________dA___________ Uro. N otebook (t page * 3 9 3 Animal No. Sex T F - r C u nc/.iL Time Dose of Cone. Dose Week I Time 24. of Hrs. Reading Time of Dose Week II Time 24 of Hrs. Reading -----1gek III : . Time of 24 Time .j. of j Dose Hrs. Reading j t ___ week IV Time Time of 24 of Dose Hrs. Readinq , 1P u Il03 He ucd <5* er cf (T v\ r-'-y VJ ABO/* O P O P / : j /!m O % : Z O A 07 / : > 0 Op 00 O i ir-/d a ;( 0 i !P 1p HC* /(r Hol 'HiFi fio H/ &O 0 ~r ___iCL P c7 f \ y / c> O T' --- ( V / 0 C-- -t ___ ^ oCL i'hi / 0 1 ^ / t O _ z ____ sm\ / :7 0 '05\.u T - p i .r- \s7kfa 0 0 0 0 0 0 0! 0< lt Xi rix 1 ~ A T > \ T~ 1( T % :i L '/at mit- <0 `W - f r A u A i r -i l a -/L ~th \ ( miA A P P/ O 7~ H 6A-^ S '- t p P ,-- Initiait A T A B L 3 (10) Du Pont HLO 432-91 Du Pont HLO 432-91 Company Sanitized. Does not certain TSCA CS! Sponsor: 07. A d i Sn I Z A/& k/i,'.:.-i Tost Article_i_J2^S_________ PHARMAKON RESEARCH INTERNATIONAL, INC. Individual Animal Scores During Induction Stu'dy Number :// yf-/y,i-/TW 9/__________ Lot Number: Notebook mo Page f} 3 W S p o n so r: - 1- O O b T e s t A r t i d e : +1 - l^^ lQ o_______________ PHARMAKON RESEARCH INTERNATIONAL, INC. In d iv id u a l Anim al Scores D uring In d u c tio n Study Number jYt-fxn -9/ fy M n w M / N1 ote b ook L o t Number: ______ d_________ Page ft N70 Anim al No. Sex Dose Cone. T im e of Dose Week X T im e 24 o f H rs. Reading T im e of Dose Week I I T im e 24 o f H rs. Reading . Tim of e Week 24 I I I T im e of Dose H rs. Reading Tim e of! Week iv 24 Tiomf e Dose ' H rs. Reading c. (Y/M -i L L : Imi /.W /* O 11:o,D litt & F7 ----- O <o I t ? 1 f f A5--N^O 1 )o r Z . rD $ W / iV u n :& > 2. & 0 1 _ J ____ !/ I f .' I f 9 0 _ / / o //.- H A * UDh C' 1 1 / F ri Ln U9 3 llCFl 1 O o 1 / eO / d rh II,'J S h .'i; 11fi 0 ~or -v \r /mm K j-W * - oI I \7~ \ / / n / 3.s* ?:/ / 7 ' \/ %; 3 7 S~7 *'# r J . ____ N/ \ / \J/ //:n w ~?oF7l,-/9// cH/Ao/tUe. f/MM rA% l / i-cF1 \lF /fates C": Du Pont HLO 432-91 In itia it Ehi =<tdfma_ EY A TA PL3(10) _!_L Du Pont HLO 432-91 Company Sanitized. Does not contain TSC, ATABL3(10) r CV W n PHARMAKON RESEARi_n IN TERN ATIO N AL, IN C . 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Doss not contain TSC.A CB l I n itia ls ATAHL3(10) O'* o & < zu A .*1 cn PHARMAKON RESEARun INTERNATIONAL, INC. Sponsor: Test A rtic le Individual Animal Scores after Challenge ,9 / M 0 U /s Study Number : P U 0 0 / ~ 9 / A ______ Lot Numbers____ fO / / _____ N7Notebook # oQ/j - ^a9e #---- t^ L Animal No. Sex Dose Dose Cone. Cone. / Time Left Right / of Shoulder Shouldqr Dose f1t /0f1 0-7 //ao y r , n a 3 f(0 //flr r t f " n tu T \ {ntQl 3SJ. - T l/OQ h V^ -- / / / / 1 1 1 //.VO |t HQC 1 \/ 1 fJ Challenge Time Time 24 of 48 of Hrs. Reading Hrs. Reading ir.lO fir* ?. //0/ i 2V? 3v / / 3 7/ ib / / / \Jz' 0 //.O l A" ) -- (oh) c<6---- " 3? y v Dose Cone. Left Shoulder Dose Cone. Time Right of Shoulder Dose Rechallenge 7 Time T^me 24 of 48 / o f Hrs Reading Hrsv Reading / -- / / / / ,/ / ______J. / ______/ / / Initia)Ls / ATABL3 (10) Du Pont HLO 432-91